Manipulation of cytokinin level in the ergot fungus Claviceps purpurea emphasizes its contribution to virulence

Abstract

Pathogen-derived cytokinins (CKs) have been recognized as important virulence factor in several host–pathogen interactions and it was demonstrated multiple times that phytopathogenic fungi form CKs via the tRNA degradation pathway. In contrast to previous studies, the focus of this study is on the second step of CK formation and CK degradation to improve our understanding of the biosynthesis in fungi on the one hand, and to understand CK contribution to the infection process of Claviceps purpurea on the other hand. The ergot fungus Claviceps purpurea is a biotrophic phytopathogen with a broad host range including economically important crops causing harvest intoxication upon infection. Its infection process is restricted to unfertilized ovaries without causing macroscopic defense symptoms. Thus, sophisticated host manipulation strategies are implicated. The cytokinin (CK) plant hormones are known to regulate diverse plant cell processes, and several plant pathogens alter CK levels during infection. C. purpurea synthesizes CKs via two mechanisms, and fungus-derived CKs influence the host–pathogen interaction but not fungus itself. CK deficiency in fungi with impact on virulence has only been achieved to date by deletion of a tRNA-ipt gene that is also involved in a process of translation regulation. To obtain a better understanding of CK biosynthesis and CKs’ contribution to the plant–fungus interaction, we applied multiple approaches to generate strains with altered or depleted CK content. The first approach is based on deletion of the two CK phosphoribohydrolase (LOG)-encoding genes, which are believed to be essential for the release of active CKs. Single and double deletion strains were able to produce all types of CKs. Apparently, log gene products are dispensable for the formation of CKs and so alternative activation pathways must be present. The CK biosynthesis pathway remains unaffected in the second approach, because it is based on heterologous overexpression of CK-degrading enzymes from maize (ZmCKX1). Zmckx1 overexpressing C. purpurea strains shows strong CKX activity and drastically reduced CK levels. The strains are impaired in virulence, which reinforces the assumption that fungal-derived CKs are crucial for full virulence. Taken together, this study comprises the first analysis of a log depletion mutant that proved the presence of alternative cytokinin activation pathways in fungi and showed that heterologous CKX expression is a suitable approach for CK level reduction.

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References

  1. Akiyoshi DE, Klee H, Amasino RM et al (1984) T-DNA of Agrobacterium tumefaciens encodes an enzyme of cytokinin biosynthesis. Proc Natl Acad Sci USA 81:5994–5998

    CAS  Article  Google Scholar 

  2. Albrecht T, Argueso CT (2016) Should I fight or should I grow now? The role of cytokinins in plant growth and immunity and in the growth–defence trade-off. Ann Bot 119:725–735. https://doi.org/10.1093/aob/mcw211

    Article  PubMed Central  Google Scholar 

  3. Altschul SF, Madden TL, Schäffer AA et al (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25:3389–3402. https://doi.org/10.1093/nar/25.17.3389

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  4. Argueso CT, Ferreira FJ, Epple P, To JPC, Hutchison CE, Schaller GE, Dangl JL, Kieber JJ, McDowell JM (2012) Two-component elements mediate interactions between cytokinin and salicylic acid in plant immunity. PLoS Genetics 8:e1002448. https://doi.org/10.1371/journal.pgen.1002448

    CAS  Article  Google Scholar 

  5. Ausubel FM (1987) Current protocols in molecular biology. Wiley, New York

    Google Scholar 

  6. Bajguz A, Piotrowska A (2009) Conjugates of auxin and cytokinin. Phytochem 70:957–969. https://doi.org/10.1016/j.phytochem.2009.05.006

    CAS  Article  Google Scholar 

  7. Behr M, Motyka V, Weihmann F et al (2012) Remodeling of cytokinin metabolism at infection sites of Colletotrichum graminicola on maize leaves. Mol Plant Microbe Interact 25:1073–1082. https://doi.org/10.1094/MPMI-01-12-0012-R

    CAS  Article  PubMed  Google Scholar 

  8. Bilyeu KD, Cole JL, Laskey JG et al (2001) Molecular and biochemical characterization of a cytokinin oxidase from maize. Plant Physiol 125:378–386

    CAS  Article  Google Scholar 

  9. Boguta M, Hunter LA, Shen WC et al (1994) Subcellular locations of MOD5 proteins: mapping of sequences sufficient for targeting to mitochondria and demonstration that mitochondrial and nuclear isoforms commingle in the cytosol. Mol Cell Biol 14:2298–2306

    CAS  Article  Google Scholar 

  10. Bruce SA, Saville BJ, Neil Emery RJ (2011) Ustilago maydis produces cytokinins and abscisic acid for potential regulation of tumor formation in maize. J Plant Growth Regul 30:51–63. https://doi.org/10.1007/s00344-010-9166-8

    CAS  Article  Google Scholar 

  11. Carimi F, Zottini M, Formentin E et al (2003) Cytokinins: new apoptotic inducers in plants. Planta 216:413–421. https://doi.org/10.1007/s00425-002-0862-x

    CAS  Article  PubMed  Google Scholar 

  12. Cenis JL (1992) Rapid extraction of fungal DNA for PCR amplification. Nucleic Acids Res 20:2380

    CAS  Article  Google Scholar 

  13. Cesari S, Thilliez G, Ribot C et al (2013) The rice resistance protein pair RGA4/RGA5 recognizes the Magnaporthe oryzae effectors AVR-Pia and AVR1-CO39 by direct binding. Plant Cell 25:1463–1481. https://doi.org/10.1105/tpc.112.107201

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  14. Chanclud E, Morel J-B (2016) Plant hormones: a fungal point of view. Mol Plant Pathol 17:1289–1297. https://doi.org/10.1111/mpp.12393

    Article  PubMed  Google Scholar 

  15. Chanclud E, Kisiala A, Emery NRJ et al (2016) Cytokinin production by the rice blast fungus is a pivotal requirement for full virulence. PLoS Pathog 12:e1005457. https://doi.org/10.1371/journal.ppat.1005457

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  16. Chen C (1997) Cytokinin biosynthesis and interconversion. Physiol Plant 101:665–673. https://doi.org/10.1111/j.1399-3054.1997.tb01051.x

    CAS  Article  Google Scholar 

  17. Christianson TW, Sikorski RS, Dante M et al (1992) Multifunctional yeast high-copy-number shuttle vectors. Gene 110:119–122. https://doi.org/10.1016/0378-1119(92)90454-W

    CAS  Article  PubMed  Google Scholar 

  18. Colot HV, Park G, Turner GE et al (2006) A high-throughput gene knockout procedure for Neurospora reveals functions for multiple transcription factors. Proc Natl Acad Sci USA 103:10352–10357. https://doi.org/10.1073/pnas.0601456103

    CAS  Article  PubMed  Google Scholar 

  19. Crespi M, Messens E, Caplan AB et al (1992) Fasciation induction by the phytopathogen Rhodococcus fascians depends upon a linear plasmid encoding a cytokinin synthase gene. EMBO J 11:795–804

    CAS  Article  Google Scholar 

  20. Dihanich ME, Najarian D, Clark R et al (1987) Isolation and characterization of MOD5, a gene required for isopentenylation of cytoplasmic and mitochondrial tRNAs of Saccharomyces cerevisiae. Mol Cell Biol 7:177–184. https://doi.org/10.1128/MCB.7.1.177

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  21. Donaton MCV, Holsbeeks I, Lagatie O et al (2003) The Gap1 general amino acid permease acts as an amino acid sensor for activation of protein kinase A targets in the yeast Saccharomyces cerevisiae. Mol Microbiol 50:911–929. https://doi.org/10.1046/j.1365-2958.2003.03732.x

    CAS  Article  PubMed  Google Scholar 

  22. Dzurová L, Forneris F, Savino S et al (2015) The three-dimensional structure of ‘Lonely Guy’ from Claviceps purpurea provides insights into the phosphoribohydrolase function of Rossmann fold-containing lysine decarboxylase-like proteins. Proteins Struct Funct Bioinform 83:1539–1546. https://doi.org/10.1002/prot.24835

    CAS  Article  Google Scholar 

  23. El Yacoubi B, Bailly M, de Crécy-Lagard V (2012) Biosynthesis and function of posttranscriptional modifications of transfer RNAs. Annu Rev Genet 46:69–95. https://doi.org/10.1146/annurev-genet-110711-155641

    CAS  Article  PubMed  Google Scholar 

  24. Esser K, Tudzynski P (1978) Genetics of the ergot fungus Claviceps purpurea. Theor Appl Genet 53:145–149. https://doi.org/10.1007/BF00273574

    CAS  Article  PubMed  Google Scholar 

  25. Frébort I, Šebela M, Galuszka P et al (2002) Cytokinin oxidase/cytokinin dehydrogenase assay: optimized procedures and applications. Anal Biochem 306:1–7. https://doi.org/10.1006/abio.2002.5670

    CAS  Article  PubMed  Google Scholar 

  26. Galuszka P, Spíchal L, Kopečný D et al (2008) Metabolism of plant hormones cytokinins and their function in signaling, cell differentiation and plant development. In: Atta-ur-Rahman (ed) Studies in Natural Products Chemistry, vol 34. Elsevier, Amsterdam, 203–264

    Google Scholar 

  27. Gan S, Amasino RM (1995) Inhibition of leaf senescence by autoregulated production of cytokinin. Science 270:1986–1988. https://doi.org/10.1126/science.270.5244.1986

    CAS  Article  PubMed  Google Scholar 

  28. Gardiner DM, Jarvis RS, Howlett BJ (2005) The ABC transporter gene in the sirodesmin biosynthetic gene cluster of Leptosphaeria maculans is not essential for sirodesmin production but facilitates self-protection. Fungal Genet Biol 42:257–263. https://doi.org/10.1016/j.fgb.2004.12.001

    CAS  Article  PubMed  Google Scholar 

  29. Goble AM, Fan H, Sali A, Raushel FM (2011) Discovery of a cytokinin deaminase. ACS Chem Biol 6:1036–1040. https://doi.org/10.1021/cb200198c

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  30. Golovko A, Sitbon F, Tillberg E, Nicander B (2002) Identification of a tRNA isopentenyltransferase gene from Arabidopsis thaliana. Plant Mol Biol 49:161–169

    CAS  Article  Google Scholar 

  31. Hann DR, Domínguez-Ferreras A, Motyka V et al (2014) The Pseudomonas type III effector HopQ1 activates cytokinin signaling and interferes with plant innate immunity. New Phytol 201:585–598. https://doi.org/10.1111/nph.12544

    CAS  Article  PubMed  Google Scholar 

  32. Hérivaux A, Dugé de Bernonville T, Roux C et al (2017) The identification of phytohormone receptor homologs in early diverging fungi suggests a role for plant sensing in land colonization by fungi. MBio 8:e01739-16. https://doi.org/10.1128/mBio.01739-16

    Article  PubMed  PubMed Central  Google Scholar 

  33. Hinsch J, Tudzynski P (2015) Claviceps: the Ergot fungus. In: Paterson R, Lima N (eds) Molecular biology of food and water borne mycotoxigenic and mycotic fungi, 1st edn. CRC Press, Boca Raton, 229–250

    Google Scholar 

  34. Hinsch J, Vrabka J, Oeser B et al (2015) De novo biosynthesis of cytokinins in the biotrophic fungus Claviceps purpurea. Environ Microbiol 17:2935–2951. https://doi.org/10.1111/1462-2920.12838

    CAS  Article  PubMed  Google Scholar 

  35. Hinsch J, Galuszka P, Tudzynski P (2016) Functional characterization of the first filamentous fungal tRNA-isopentenyltransferase and its role in the virulence of Claviceps purpurea. New Phytol 211:980–992. https://doi.org/10.1111/nph.13960

    CAS  Article  PubMed  Google Scholar 

  36. Hluska T, Dobrev PI, Tarkowská D et al (2016) Cytokinin metabolism in maize: Novel evidence of cytokinin abundance, interconversions and formation of a new trans-zeatin metabolic product with a weak anticytokinin activity. Plant Sci 247:127–137. https://doi.org/10.1016/j.plantsci.2016.03.014

    CAS  Article  PubMed  Google Scholar 

  37. Huang S, Cerny RE, Qi Y et al (2003) Transgenic studies on the involvement of cytokinin and gibberellin in male development. Plant Physiol 131:1270–1282. https://doi.org/10.1104/pp.102.018598

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  38. Hüsgen U, Buttner P, Muller U, Tudzynski P (1999) Variation in karyotype and ploidy level among field isolates of Claviceps purpurea. J Phytopathol 147:591–597. https://doi.org/10.1046/j.1439-0434.1999.00432.x

    Article  Google Scholar 

  39. Janevska S, Arndt B, Niehaus E-M et al (2016) Gibepyrone biosynthesis in the rice pathogen Fusarium fujikuroi is facilitated by a small polyketide synthase gene cluster. J Biol Chem 291:27403–27420. https://doi.org/10.1074/jbc.M116.753053

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  40. Jiang C-J, Shimono M, Sugano S et al (2013) Cytokinins act synergistically with salicylic acid to activate defense gene expression in rice. Mol Plant Microbe Interact 26:287–296. https://doi.org/10.1094/MPMI-06-12-0152-R

    CAS  Article  PubMed  Google Scholar 

  41. Jiang Y, Jiang Q, Hao C et al (2015) A yield-associated gene TaCWI, in wheat: its function, selection and evolution in global breeding revealed by haplotype analysis. Theor Appl Genet 128:131–143. https://doi.org/10.1007/s00122-014-2417-5

    CAS  Article  PubMed  Google Scholar 

  42. Jungehülsing U, Arntz C, Smit R, Tudzynski P (1994) The Claviceps purpurea glyceraldehyde-3-phosphate dehydrogenase gene: cloning, characterization, and use for the improvement of a dominant selection system. Curr Genet 25:101–106. https://doi.org/10.1007/BF00309533

    Article  PubMed  Google Scholar 

  43. Kim D, Pertea G, Trapnell C et al (2013) TopHat2: accurate alignment of transcriptomes in the presence of insertions, deletions and gene fusions. Genome Biol 14:R36. https://doi.org/10.1186/gb-2013-14-4-r36

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  44. Konevega AL, Soboleva NG, Makhno VI et al (2005) The effect of modification of tRNA nucleotide-37 on the tRNA interaction with the P- and A-site of the 70S ribosome Escherichia coli. Mol Biol (Mosk) 40:669–683

    Google Scholar 

  45. Kopečná M, Blaschke H, Kopečný D et al (2013) Structure and function of nucleoside hydrolases from Physcomitrella patens and maize catalyzing the hydrolysis of purine, pyrimidine, and cytokinin ribosides. Plant Physiol 163:1568–1583. https://doi.org/10.1104/pp.113.228775

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  46. Kopečný D, Pethe C, Šebela M et al (2005) High-level expression and characterization of Zea mays cytokinin oxidase/dehydrogenase in Yarrowia lipolytica. Biochimie 87:1011–1022. https://doi.org/10.1016/j.biochi.2005.04.006

    CAS  Article  PubMed  Google Scholar 

  47. Kurakawa T, Ueda N, Maekawa M et al (2007) Direct control of shoot meristem activity by a cytokinin-activating enzyme. Nature 445:652–655. https://doi.org/10.1038/nature05504

    CAS  Article  PubMed  Google Scholar 

  48. Lamichhane TN, Blewett NH, Crawford AK et al (2013) Lack of tRNA modification isopentenyl-A37 alters mRNA decoding and causes metabolic deficiencies in fission yeast. Mol Cell Biol 33:2918–2929. https://doi.org/10.1128/MCB.00278-13

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  49. Lara MEB, Garcia M-CG, Fatima T et al (2004) Extracellular invertase is an essential component of cytokinin-mediated delay of senescence. Plant Cell 16:1276–1287. https://doi.org/10.1105/tpc.018929

    Article  Google Scholar 

  50. Liao Y, Smyth GK, Shi W (2014) FeatureCounts: an efficient general purpose program for assigning sequence reads to genomic features. Bioinformatics 30:923–930. https://doi.org/10.1093/bioinformatics/btt656

    CAS  Article  Google Scholar 

  51. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2−∆∆CT method. Methods 25:402–408. https://doi.org/10.1006/meth.2001.1262

    CAS  Article  PubMed  Google Scholar 

  52. Lomin SN, Krivosheev DM, Steklov MY et al (2015) Plant membrane assays with cytokinin receptors underpin the unique role of free cytokinin bases as biologically active ligands. J Exp Bot 66:1851–1863. https://doi.org/10.1093/jxb/eru522

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  53. Love MI, Huber W, Anders S (2014) Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2. Genome Biol 15:1–21. https://doi.org/10.1186/s13059-014-0550-8

    CAS  Article  Google Scholar 

  54. Mantle PG, Nisbet LJ (1976) Differentiation of Claviceps purpurea in axenic culture. J Gen Microbiol 93:321–334. https://doi.org/10.1099/00221287-93-2-321

    CAS  Article  PubMed  Google Scholar 

  55. Mitsuhara I, Iwai T, Seo S et al (2008) Characteristic expression of twelve rice PR1 family genes in response to pathogen infection, wounding, and defense-related signal compounds. Mol Genet Genom 279:415–427. https://doi.org/10.1007/s00438-008-0322-9

    CAS  Article  Google Scholar 

  56. Mlejnek P, Procházka S (2002) Activation of caspase-like proteases and induction of apoptosis by isopentenyladenosine in tobacco BY-2 cells. Planta 215:158–166. https://doi.org/10.1007/s00425-002-0733-5

    CAS  Article  PubMed  Google Scholar 

  57. Moore S, De Vries OMH, Tudzynski P (2002) The major Cu,Zn SOD of the phytopathogen Claviceps purpurea is not essential for pathogenicity. Mol Plant Pathol 3:9–22. https://doi.org/10.1046/j.1464-6722.2001.00088.x

    CAS  Article  PubMed  Google Scholar 

  58. Morrison EN, Knowles S, Hayward A et al (2015) Detection of phytohormones in temperate forest fungi predicts consistent abscisic acid production and a common pathway for cytokinin biosynthesis. Mycologia 107:245–257. https://doi.org/10.3852/14-157

    CAS  Article  PubMed  Google Scholar 

  59. Morrison EN, Emery RJN, Saville BJ (2017) Fungal derived cytokinins are necessary for normal Ustilago maydis infection of maize. Plant Pathol 66:726–742. https://doi.org/10.1111/ppa.12629

    CAS  Article  Google Scholar 

  60. Naseem M, Sarukhanyan E, Dandekar T (2015) LONELY-GUY Knocks Every Door: Crosskingdom Microbial Pathogenesis. Trends Plant Sci 20:781–783. https://doi.org/10.1016/j.tplants.2015.10.017

    CAS  Article  PubMed  Google Scholar 

  61. Niehaus E-M, Münsterkötter M, Proctor RH et al (2016) Comparative ‘Omics’ of the Fusarium fujikuroi species complex highlights differences in genetic potential and metabolite synthesis. Genome Biol Evol 8:3574–3599. https://doi.org/10.1093/gbe/evw259

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  62. Oeser B, Kind S, Schurack S et al (2017) Cross-talk of the biotrophic pathogen Claviceps purpurea and its host Secale cereale. BMC Genom 18:273. https://doi.org/10.1186/s12864-017-3619-4

    CAS  Article  Google Scholar 

  63. Persson BC, Ólafsson Ó, Lundgren HK, Björk GR (1998) The ms 2io6A37 modification of tRNA in Salmonella typhimurium regulates growth on citric acid cycle intermediates. J Bacteriol 180:3144–3151

    CAS  PubMed  PubMed Central  Google Scholar 

  64. Pertry I, Václavíková K, Depuydt S et al (2009) Identification of Rhodococcus fascians cytokinins and their modus operandi to reshape the plant. Proc Natl Acad Sci USA 106:929–934. https://doi.org/10.1073/pnas.0811683106

    CAS  Article  PubMed  Google Scholar 

  65. Phizicky EM, Hopper AK (2010) tRNA biology charges to the front. Genes Dev 24:1832–1860. https://doi.org/10.1101/gad.1956510

    Article  PubMed  PubMed Central  Google Scholar 

  66. Pospíšilová H, Šebela M, Novák O, Frébort I (2008) Hydrolytic cleavage of N6-substituted adenine derivatives by eukaryotic adenine and adenosine deaminases. Biosci Rep 28:335–347. https://doi.org/10.1042/BSR20080081

    CAS  Article  PubMed  Google Scholar 

  67. Pratt-Hyatt M, Pai DA, Haeusler RA et al (2013) Mod5 protein binds to tRNA gene complexes and affects local transcriptional silencing. Proc Natl Acad Sci USA 110:3081–3089. https://doi.org/10.1073/pnas.1219946110

    Article  Google Scholar 

  68. Roitsch T, Ehneß R (2000) Regulation of source/sink relations by cytokinins. Plant Growth Regul 32:359–367. https://doi.org/10.1023/A:1010781500705

    CAS  Article  Google Scholar 

  69. Sakakibara H (2006) Cytokinins: activity, biosynthesis, and translocation. Annu Rev Plant Biol 57:431–449. https://doi.org/10.1146/annurev.arplant.57.032905.105231

    CAS  Article  PubMed  Google Scholar 

  70. Samanovic MI, Tu S, Novák O et al (2015) Proteasomal control of cytokinin synthesis protects Mycobacterium tuberculosis against nitric oxide. Mol Cell 57:984–994. https://doi.org/10.1016/j.molcel.2015.01.024

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  71. Sambrook J, Fritsch E, Maniatis T (1989) Molecular Cloning. A laboratory manual, 2nd edn. Cold Spring Harbor Laboratory Press, New York

    Google Scholar 

  72. Schardl CL, Young CA, Hesse U et al (2013) Plant-symbiotic fungi as chemical engineers: multi-genome analysis of the clavicipitaceae reveals dynamics of alkaloid loci. PLoS Genet 9:e1003323. https://doi.org/10.1371/journal.pgen.1003323

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  73. Scheffer J, Tudzynski P (2006) In vitro pathogenicity assay for the ergot fungus Claviceps purpurea. Mycol Res 110:465–470. https://doi.org/10.1016/j.mycres.2005.11.011

    Article  PubMed  Google Scholar 

  74. Schmülling T, Werner T, Riefler M et al (2003) Structure and function of cytokinin oxidase/dehydrogenase genes of maize, rice, Arabidopsis and other species. J Plant Res 116:241–252. https://doi.org/10.1007/s10265-003-0096-4

    CAS  Article  PubMed  Google Scholar 

  75. Schumacher J (2012) Tools for Botrytis cinerea: new expression vectors make the gray mold fungus more accessible to cell biology approaches. Fungal Genet Biol 49:483–497. https://doi.org/10.1016/j.fgb.2012.03.005

    CAS  Article  PubMed  Google Scholar 

  76. Shigenaga AM, Argueso CT (2016) No hormone to rule them all: interactions of plant hormones during the responses of plants to pathogens. Semin Cell Dev Biol 56:174–189. https://doi.org/10.1016/j.semcdb.2016.06.005

    CAS  Article  PubMed  Google Scholar 

  77. Sørensen JL, Benfield AH, Wollenberg RD et al (2017) The cereal pathogen Fusarium pseudograminearum produces a new class of active cytokinins during infection. Mol Plant Pathol. https://doi.org/10.1111/mpp.12593 (in press)

    Article  PubMed  Google Scholar 

  78. Suzuki G, Shimazu N, Tanaka M (2012) A yeast prion, Mod5, promotes acquired drug resistance and cell survival under environmental stress. Science 336:355–359. https://doi.org/10.1126/science.1219491

    CAS  Article  PubMed  Google Scholar 

  79. Takei K, Yamaya T, Sakakibara H (2004) Arabidopsis CYP735A1 and CYP735A2 encode cytokinin hydroxylases that catalyze the biosynthesis of trans-zeatin. J Biol Chem 279:41866–41872. https://doi.org/10.1074/jbc.M406337200

    CAS  Article  PubMed  Google Scholar 

  80. Tenberge K, Homman V, Oeser B, Tudzynski P (1996) Structure and expression of two polygalacturonase genes of Claviceps purpurea oriented in tandem and cytological evidence for pectinolytic enzyme activity during infection of rye. Phytopathology 86:1084–1097. https://doi.org/10.1094/Phyto-86-1084

    CAS  Article  Google Scholar 

  81. Trdá L, Barešová M, Šašek V et al (2017) Cytokinin metabolism of pathogenic fungus Leptosphaeria maculans involves isopentenyltransferase, adenosine kinase and cytokinin oxidase/dehydrogenase. Front Microbiol 8:1374. https://doi.org/10.3389/fmicb.2017.01374

    Article  PubMed  PubMed Central  Google Scholar 

  82. Walters DR, McRoberts N (2006) Plants and biotrophs: a pivotal role for cytokinins? Trends Plant Sci 11:581–586. https://doi.org/10.1016/j.tplants.2006.10.003

    CAS  Article  PubMed  Google Scholar 

  83. Werner T, Motyka V, Laucou V et al (2003) Cytokinin-deficient transgenic Arabidopsis plants show multiple developmental alterations indicating opposite functions of cytokinins in the regulation of shoot and root meristem activity. Plant Cell 15:2532–2550. https://doi.org/10.1105/tpc.014928

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  84. Winston F, Dollard C, Ricupero-Hovasse SL (1995) Construction of a set of convenient Saccharomyces cerevisiae strains that are isogenic to S288C. Yeast 11:53–55. https://doi.org/10.1002/yea.320110107

    CAS  Article  PubMed  Google Scholar 

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Acknowledgements

We thank Daniela Odinius for excellent technical assistance. This study was supported by grants from the International Graduate School (GRK1409), Sino-German-Science Center (GZ928), the National Science Foundation, Czech Republic (Grant No. 16-10602S) and the National Program of Sustainability I, Ministry of Education, Youth and Sports, Czech Republic (Grant No. LO1204).

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Communicated by M. Kupiec.

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Kind, S., Hinsch, J., Vrabka, J. et al. Manipulation of cytokinin level in the ergot fungus Claviceps purpurea emphasizes its contribution to virulence. Curr Genet 64, 1303–1319 (2018). https://doi.org/10.1007/s00294-018-0847-3

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Keywords

  • Claviceps purpurea
  • Cytokinin
  • Cytokinin oxidase/dehydrogenase
  • Phosphoribohydrolase
  • Virulence